BioMaster UDG HS-qPCR Lo-ROX SYBR (2×)

Features
2× reaction mix includes Hot-Start Taq DNA polymerase, dUTP and N-uracil-DNA glycosylase for real-time PCR with SYBR Green I; contains ROX at low concentration for studies involving use of specific equipment.
Application
  • Hot-start real-time PCR with oligonucleotide probes
  • Conventional PCR
  • High-throughput PCR
  • Genotyping

Basic description

BioMaster UDG HS-qPCR Lo-ROX SYBR (2×) contains 2× BioMaster UDG HS-qPCR Lo-ROX SYBR reaction mix, 50 mM MgCl2 solution and sterile water. The mix is developed for quantitative real-time PCR with fluorescent dye SYBR Green I on PCR platforms that utilize ROX as a reference dye. BioMaster UDG HS-qPCR Lo-ROX SYBR (2×) includes all components (except for DNA template and primers) necessary for PCR: highly processive recombinant HS-Taq DNA polymerase, N-uracil-DNA glycosylase, dNTP mix, PCR buffer, Mg2+ solution, SYBR Green I, fluorescent dye ROX, and inert dye.

BioMaster UDG HS-qPCR Lo-ROX SYBR (2×) is optimized for efficient and reproducible hot-start real-time PCR of genomic, plasmid and viral DNA samples. BioMaster UDG HS-qPCR Lo-ROX SYBR (2×) mix contains substances influencing primer annealing temperature and characteristics of template melting thus enabling to increase PCR specificity and use templates with complicated spatial structure.

N-uracil-DNA glycosylase (UDG) and dUTP (in proportion with dTTP) provide protection from amplicon transfer between reaction solutions (cross-contamination). DNA polymerase included in the mix is inactive at room temperature, its activation requires preheating at 95 °C for 5 min. Blue hue of the reaction solution provided by the inert dye allows control when using multi-well plates. The master mix is ideally suitable for PCR platforms that use ROX passive dye as a reference guide, such as Life Technologies (ABI) 7500, 7500 Fast, ViiA 7, QuantStudio 12K; Stratagene Mx4000, Mx3005P, and Mx3000P.

Areas of use

  • Classic PCR with "hot" start
  • Genotyping

Reaction mixture properties

  • The mix is inactive at room temperature due to the Hot-Start technology and is activated after incubation at 95 °C for 5 min
  • Allows normalization using ROX dye as a reference
  • The presence of dUTP ensures incorporation of uridine into each synthesized DNA strand, UDG is capable of eliminating uracil from single- and double-stranded DNA molecules
  • The mix is optimized for the specific performance of HS-Taq DNA polymerase, long-term storage (the storage of BioMaster UDG HS-qPCR Lo-ROX SYBR (2×) at room temperature for 7 days does not affect PCR efficiency), multiple thawing-freezing cycles

Benefits of using

  • Enzyme with hot-start capability increases reaction specificity
  • Activation of HS-Taq DNA polymerase requires not more than 5 min heating
  • High selectivity and reaction yield
  • Reduced preparation time
  • Prevents re-amplification of PCR carryover products
  • Allows data normalization
  • Standardized conditions of the same-type reactions (reduced pipetting error during mixing PCR components from experiment to experiment)
  • Minimized efforts

Storage

1 year (under proper storage and transportation conditions).

Transportation

at -20 °С; not more than 50 thawing-freezing cycles.

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